WO2020109801A1 - Dispositifs, systèmes et procédés microfluidiques améliorés - Google Patents

Dispositifs, systèmes et procédés microfluidiques améliorés Download PDF

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Publication number
WO2020109801A1
WO2020109801A1 PCT/GB2019/053367 GB2019053367W WO2020109801A1 WO 2020109801 A1 WO2020109801 A1 WO 2020109801A1 GB 2019053367 W GB2019053367 W GB 2019053367W WO 2020109801 A1 WO2020109801 A1 WO 2020109801A1
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WIPO (PCT)
Prior art keywords
pressure
increase
microfluidic device
fluidically
pcr
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Ceased
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PCT/GB2019/053367
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English (en)
Inventor
Hojjat MADADI
Thomas Michael Willshare
Robin Page
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QuantumDx Group Ltd
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QuantumDx Group Ltd
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Publication date
Application filed by QuantumDx Group Ltd filed Critical QuantumDx Group Ltd
Priority to EP19816422.0A priority Critical patent/EP3887047A1/fr
Priority to US17/298,230 priority patent/US20220088590A1/en
Priority to JP2021531009A priority patent/JP7499247B2/ja
Priority to CN201980078467.9A priority patent/CN113164955B/zh
Publication of WO2020109801A1 publication Critical patent/WO2020109801A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/50273—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the means or forces applied to move the fluids
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502715—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502738—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by integrated valves
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00—Heating or cooling apparatus; Heat insulating devices
    • B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00—Heating or cooling apparatus; Heat insulating devices
    • B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
    • B01L7/525—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples with physical movement of samples between temperature zones
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/686—Polymerase chain reaction [PCR]
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0689—Sealing
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0816—Cards, e.g. flat sample carriers usually with flow in two horizontal directions
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/0883—Serpentine channels
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • B01L2400/0478—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure pistons
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • B01L2400/0481—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure squeezing of channels or chambers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • B01L2400/0487—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/06—Valves, specific forms thereof
    • B01L2400/0605—Valves, specific forms thereof check valves

Definitions

  • the present invention relates to microfluidic devices, such as cassettes, for diagnostics, which allow for on-cassette processing including amplification by polymerase chain reaction (PCR), and systems and methods of using the same, wherein microfluidic cassette is pressurised prior to amplification of nucleic acid in said sample in an amplification/PCR section of the device.
  • microfluidic devices such as cassettes, for diagnostics, which allow for on-cassette processing including amplification by polymerase chain reaction (PCR), and systems and methods of using the same, wherein microfluidic cassette is pressurised prior to amplification of nucleic acid in said sample in an amplification/PCR section of the device.
  • PCR polymerase chain reaction
  • microfluidic devices such as cassettes, that are able to perform various chemical and biochemical analyses and syntheses on device.
  • the goal is to provide devices that allow previously laboratory-based activities to be performed at the point of care (POC) in a timely and efficient manner.
  • POC point of care
  • Such microfluidic devices can be adapted for use with automated systems, thereby providing the additional benefits such as cost reductions and decreased operator errors.
  • microfluidic devices are capable of carrying out molecular techniques including the amplification of nucleic acids.
  • Amplification of DNA by polymerase chain reaction (PCR) requires reaction mixtures be subjected to repeated rounds of heating and cooling (thermocycling) whist travelling through microfluidic channels present on the cassette, which can include holding the reaction mixtures in one or more static chambers present on the cassette.
  • the temperature of the reaction mixture within the cassette therefore must be varied during a PCR cycle, and in fact varied many times during a PCR experiment, which typically requires a relatively high number of cycles to obtain a suitable amplification of the nucleic acid from the sample.
  • the variation of the temperature within the cassette does bring technical challenges.
  • denaturation of DNA typically takes place at greater than 90 degrees, and often close to 98 degrees C.
  • the temperature then needs to drop as annealing a primer to the denatured DNA is typically performed at around 45 degrees to 70 degrees C.
  • the temperature is raised again as the step of extending the annealed primers with a polymerase is typically performed at around 70 degrees to 75 degrees C.
  • Various mechanisms to allow for this have been described, with systems employing heated zones being utilised, or means for rapidly changing the temperature within microfluidic channels being employed. Such devices and systems are known.
  • 'Shuttle flow' or 'Shuttle PCR' refers to techniques in which thermocycling is performed by shuttling small plugs of PCR mixture back and forth between temperature zones. In this way temperature variations occur spatially.
  • microfluidic means with at least one dimension less than 1 millimetre and/or able to deal with microlitre or less portions of fluid.
  • cassette or “chip” means an assembled unit comprising one or more substrates with channels or chambers therein through which fluid can flow.
  • cassettes may include different regions or zones in which activities such as sample mixing, filtering, PCR amplification, identification and/or visualisation can occur and may include on-board reagents.
  • the cassettes are typically designed to be received by a diagnostic instrument such as a point-of-care (POC) instrument which incorporates additional functionality to allow a diagnostic test, or part of such a test, to be automated.
  • POC point-of-care
  • a microfluidic device comprising;
  • PCR polymerase chain reaction
  • a fluidically isolatable portion of the device comprising at least the portion of the fluidic channel having a PCR section
  • At least one closure means actuatable to fluidically isolate the fluidically isolatable portion from the external environment
  • a PCR section of the channel is a portion of the channel that has been configured to allow thermocycling of a sample passing therethrough to occur.
  • the PCR section of the channel is specifically adapted to allow a liquid sample travelling therethrough to be heated and cooled in a cyclical manner to temperatures that allow for denaturing of DNA, annealing of DNA and amplification of DNA.
  • the PCR section of the channel is heatable either by an integral heat source within the channel or the wall of the device, or more typically by being brought into the proximity of an external heat source which applies heat to microfluidic device (such as a microfluidic cassette) in a manner that allows heat to transfer into the PCR section of the channel.
  • the at least one means to increase the pressure is adapted to increase pressure within the PCR section of the channel prior to amplification of a sample by thermocycling within the PCR section.
  • the microfluidic device is suitable for receiving a liquid sample.
  • a microfluidic cassette for insertion into a point of care (POC) diagnostic instrument.
  • POC point of care
  • the fluidic channel is for transporting said liquid sample and has an inlet end and at least one second end that are in fluid communication with each other.
  • the fluidic channel is a microfluidic channel. Most preferably the circumference of the channel is fluid-tight.
  • the term 'fluid-tight' means a sealing that prevents the passage of liquids, such as water, and/or gases, in particular air, even if put under pressure (within predetermined limits). For example, the seal will remain intact up to approximately 2 bar of pressure difference between the interior and exterior sides of the slide fastener.
  • At least part of the microchannel has a serpentine configuration.
  • serpentine configurations can allow long fluid flow in a relatively small footprint, which means an efficient heat transfer by increasing surface to volume ratio.
  • the serpentine configuration can be useful in allowing fluid to travel over multiple different temperature zones, for example different heat plates, before curving back to repeat the process.
  • the serpentine configuration can be utilised along with methods of shuttling fluid back and forth between temperature zones (i.e. shuttle flow (shuttle PCR) or
  • the isolatable portion may be the entire fluidic channel present on the device. In one embodiment, substantially all of the microfluidic channel present on a cassette is the isolatable portion. In this embodiment, this means that once a sample is inserted into the cassette i.e. inserted into the start of the microfluidic channel, the channel is then sealed, and the pressure is increased.
  • a positive pressure i.e. inducing a positive pressure within the a microfluidic channel prior to significant further activity
  • other microfluidic channels within a device such as a cassette, as well as in the PCR section helps to minimise fluid(liquid)-plug breakup i.e. break-up of the sample fluid plug rather than maintaining it as a single plug of fluid (the sample plug may incorporate additional reagents as it moves through the cassette).
  • the at least one closure means is at least one air-tight valve or sealing means actuatable to fluidically isolate the fluidically isolatable portion.
  • the means to increase pressure is a positive displacement pump.
  • the means to increase pressure may be actuated to a first position to apply a first pressure to the isolatable portion, and a second position to apply a second pressure to the isolatable portion.
  • the means to increase pressure is a bellows pump.
  • the bellows pump is a substantially hemispherical compressible material with an internal cavity.
  • the bellows pump is associated with an inlet to the microfluidic channel. When pressure is applied to the external surface of the bellows pump the material is compressed such that the internal cavity is reduced in volume and any fluid in the internal cavity is pushed.
  • the bellows pump is resiliently biased to return to an expanded position.
  • the means to increase pressure is valve associable with a syringe pump.
  • the means to increase pressure is a pneumatic pressure applicator.
  • the means to increase pressure is associable with an external pressure source.
  • the means to increase pressure is an on-cassette peristaltic pump.
  • An external pressure source e.g. mechanical, pneumatic or hydraulic source of pressure can be applied by an external host instrument.
  • the plurality of means to increase the pressure comprise a plurality of positive displacement pumps, with each pump fluidically connected to the fluidically isolatable portion of the device.
  • these are two bellows pumps.
  • the plurality of means to increase the pressure are simultaneously or concurrently actuatable to increase the pressure in the isolatable portion.
  • the means to increase the pressure are actuable to a first compression position and a second compression position.
  • the second compression position is further compressed (thus having less volume within it) than the first position.
  • the means to increase pressure is adapted to increase the pressure in the isolatable portion
  • the PCR section is heatable.
  • An external source of heat can be applied by an external instrument.
  • the means for increasing pressure in the isolatable portion induces a pressure higher than atmospheric pressure.
  • the means for increasing pressure in the isolatable portion induce a pressure of 1.6bar or higher.
  • the means for increasing pressure in the isolatable portion induce a pressure of 1.2bar or higher.
  • increasing the pressure reduces the boiling point of fluids within the isolatable portion. This can be advantageous in allowing the system to be used in a range of different location, including at high altitudes.
  • the microfluidic device comprises on-board reagents for performing at least one diagnostic assay.
  • a diagnostic system comprising the microfluidic device of the first aspect, and a host instrument able to receive said microfluidic device.
  • the host instrument comprises an interface enabled to convey pressure from said host instrument to said means for increasing pressure on said microfluidic device.
  • the host instrument comprises a pneumatic interface enabled to convey pneumatic pressure from said host instrument to said means for increasing pressure on said microfluidic device.
  • the host instrument comprises a mechanical interface enabled to convey mechanical pressure from said host instrument to said means for increasing pressure on said microfluidic device.
  • the host instrument is adapted to actuate an increased pressure in the isolatable portion of the cassette when it is isolated and prior to amplification of a sample by thermocycling within the PCR section.
  • the host instrument comprises a microprocessor.
  • the microprocessor controls the interactions between the host instrument to the microfluidic device.
  • the host instrument comprises means for heating or applying heat to at least a portion of the PCR section of the device.
  • the means for heating is one or more temperature-controlling elements configured to provide at least one temperature zone.
  • PCR polymerase chain reaction
  • the sample is inserted into the device prior to the step of fluidically isolating at least a portion of the device comprising at least the PCR section.
  • the entire device is fluidically isolated.
  • the cassette can be pre-pressurised prior to polymerase chain reaction (PCR) thermocycling occurring.
  • PCR polymerase chain reaction
  • microfluidic channel only portions of the microfluidic channel are fluidically isolated.
  • branches of the channel can be closed off, including branches to reservoirs etc, either to prevent ingress of materials detrimental to PCR reactions or to reduce the volume to which pressure is applied.
  • one or more fluid-tight valves are used to fluidically isolate the system.
  • valves may be within the channels or form the inlet (or outlet if present).
  • the step of increasing the pressure comprises actuating a plurality of means to increase the pressure.
  • the step of increasing the pressure comprises compressing a plurality of positive displacement pumps.
  • the step of increasing the pressure comprises compressing a plurality of positive displacement pumps simultaneously or concurrently to increase the pressure in the isolatable portion.
  • the step of increasing the pressure comprises compressing a plurality of positive displacement pumps to a first position, wherein the positive displacement pumps may still be further compressed to at least a second position.
  • the step of increasing the pressure increases the pressure in the isolated portion to greater than 1.2bar , more preferably to greater than 1.4bar yet more preferably greater than 1.6bar.
  • the step of increasing the pressure increases the pressure in the isolated portion to between 1.5bar and 2bar.
  • the step of increasing the pressure increases the pressure in the isolated portion to 1.6bar.
  • microfluidic means with at least one dimension less than 1 millimetre and/or able to deal with microlitre or less portions of fluid.
  • Figure la provides a perspective view of a microfluidics cassette according to an aspect of the present invention, the view showing an outer surface;
  • Figure lb provides an exploded perspective view of a similar microfluidics cassette shown from the other side, with various internal features visible.
  • Figure 2a, 2b, 2c is a simplified diagram showing the use of two bellows pumps to pressurise a sealed area of a cassette, with the first applying pressure and the second acting as a reservoir; and
  • Figure 3a, 3b, 3c is a simplified diagram showing the use of two bellows pumps that are actuated simultaneously pressurise a sealed area of a cassette.
  • Fig 3B shows the pressurization step while
  • Fig. 3C shows the reciprocation mode to move the sample while keeping the pressure in a sellable portion of the cassette.
  • a microfluidic cassette 1 more generally termed a 'device', with a micro- channel 2, where the micro-channel 2 allows for continuous flow-through of fluid as required.
  • the micro-channel 2 is formed inside the microfluidic cassette 1, in the desired length and shape to allow the passage of a sample, typically a biological sample in liquid format, and/or reagents, some of which may be incorporated on-cassette during the flow-through, along a fluid flow path and through various zones or areas which allow different activities to occur including amplification of DNA from the sample by polymerase chain reaction (PCR).
  • PCR polymerase chain reaction
  • a PCR section of the channel 7 is a portion of the channel that has been configured to allow thermocycling of a sample passing therethrough to occur.
  • the PCR section 7 of the channel (and cassette) is specifically adapted to allow a liquid sample travelling therethrough to be heated and cooled in a cyclical manner to temperatures that allow for denaturing of DNA, annealing of DNA and simplification of DNA.
  • the PCR section 7 of the channel is heatable either by an integral heat source within the channel or the wall of the device, or more typically by being brought into the proximity of an external heat source which applies heat to microfluidic device (such as a microfluidic cassette) in a manner that allows heat to transfer into the PCR section of the channel.
  • valves and fluidically intercommunicating offshoots such as additional channels, reservoirs or chambers can be used to allow mixing, washing, removal and other actions to occur according to the needs of a diagnostic or biochemical assay to be performed therein in an automated or semi-automated manner.
  • the assay is generally carried out by allowing a sample to interact and/or react with one or more reagents in one or more steps; typically in one or more channels or chambers of the device, for times and at temperatures effective in forming a detectable product that indicates the presence or absence of an analyte in the sample.
  • the channel 2 is formed in a first surface of a first substrate 3, typically a substantially planar, substantially rigid substrate which in this embodiment is polypropylene.
  • the first substrate 3 is overlaid with a second substrate 4, which in this embodiment is a polypropylene film.
  • a substantially closed channel 2 is provided (inlets and outlets can be included as required).
  • the first substrate 3 is a planar element with an upper and lower surface, the majority of the microchannel 2 can formed in the upper surface or the lower surface.
  • the second substrate i.e. the film 4 forms the upper wall of the microchannel 2 in use.
  • the use of laser welding ensured that a fluid-tight, and more specifically an air-tight seal is created around the microchannels, such that no air can escape or leak out between the first and second substrates.
  • the seal or weld is sufficiently strong to withstand increased pressure levels within the cassette compare to outside of the cassette. Typically, it is strong enough to withstand internal gauge pressures of at least 2bar.
  • this embodiment has the second substrate as a film 4, the second substrate can be another material and may itself have grooves or channel formed on its surface that can be aligned with the channels of the first substrate.
  • a closed channel 2 is provided (again inlets and outlets can be included as required).
  • the cartridges are typically consumables; i.e., they are used once and then discarded; and contain all or many of the reagents needed for one or more assays to be performed. Such reagents can be held on the cartridge in reservoirs or similar. Where necessary, the first and second substrates 3, 4 can be aligned prior to bonding.
  • the length and cross-sectional shape of the channel 2 can be any appropriate shape to allow for the desired transport and processing of a sample and or reagents.
  • Such microfluidic systems 'lab-on-a-chip' type systems being well known in the art.
  • the cassette 1 is provided with an inlet 5 for receiving a sample into the microfluidic channel.
  • the inlet 5 is provided with an air-tight seal in the form of an insertable cap 6.
  • the insertable cap 6 is hinged to the surface of the cassette for ease of use.
  • the cap 6 is sized to be received into the aperture of the inlet and is provided with a resiliently deformable collar that acts to form an air-tight seal in the inlet after the sample has been inserted and when the cap 6 is closed.
  • valves positioned within the microchannel, away from the inlet can be used to create a smaller volume air-tight area if required (still containing the PCR amplification zone, or PCR section 7).
  • a first air-tight valve could be provided proximate and upstream of the amplification zone 7 and a second air-tight valve could be provided proximate and upstream of the amplification zone 7.
  • the microchannel 2 is a continuous flow channel and, in this embodiment, it includes an amplification zone or PCR section 7 which includes three portions of channel that are heatable to different temperatures to allow the thermocycling of temperatures.
  • a further portion of the channel in the PCR section 7 is heated to a temperature of around 70 degrees to 75 degrees C.
  • the heating of the channel occurs when an external heater source is brought into close proximity with the cassette 1 such that heat exchange occurs through the wall of the cassette into the relevant portions of the micro-channel.
  • the microchannel in the amplification zone or PCR section 7 is serpentine in shape such that the sample will travel through the different temperature zones as it travels along each section of the serpentine shape.
  • the heaters are provided in a separate host instrument which receives the cassette therein, but in theory the heaters could form part of the cassette, for example being positioned within the channels themselves.
  • a serpentine shaped microchannel 2 is used in the PCR section 7 to ensure sample moving through the channel is cyclically taken through the different temperatures required for a PCR reaction to occur
  • a shuttle flow system could be used in place of the serpentine shaping.
  • the sample is shuttled back and forward between two or more sections of the microchannel (as opposed to continuously moving forward as in the serpentine variant), each section being held at one of the required temperatures for a PCR reaction to occur.
  • the microchannel includes a first bellows pump 8A in fluid communication with the microchannel 2 (via bellows inlet 10A).
  • the first bellows pump 8A is a positive displacement pump, comprising a hemispherical, compressible bellow which is resiliently biased to return to its expanded shape.
  • the internal dimensions of the hemispherical bellow can be changed by applying pressure to its external surface c.
  • mechanical actuators found on a host instrument into which the cassette is placed act under the control of a microprocessor to actuate the bellows pumps by compression, partial compression, partial decompression or decompression of the same.
  • the actuators could however, in theory, be incorporated onto the cassette.
  • the microchannel also includes a second bellows pump 8B in fluid communication with the microchannel 2 (via bellows inlet 10B).
  • the second bellows pump 8B is also a positive displacement pump, in this example again comprising a hemispherical, compressible bellow which is resiliently biased to return to its expanded shape.
  • the second bellows 8B can be partially compressed or decompressed as well as fully compressed or decompressed.
  • the microchannel 2 also includes various other reservoirs, branches and chambers (such as a capture and viewing chamber for example) as are required to carry out a molecular assay.
  • the host instrument will include elements such as actuators, heaters and optical elements.
  • bellows pumps with a hemispherical bellow
  • other bellows formations could be used, or indeed alternative types of positive displacement pump could be used.
  • another type of positive displacement pump is a syringe pump in which the displacement of the piston of a syringe inside a syringe cylinder causes fluid to be sucked into or pressed out of the syringe outlet.
  • a liquid sample 9 is introduced into the inlet 5, which leads to the micro-fluidic channel 2 and the cap 6 is closed to form an air-tight seal.
  • the cassette 1 is placed into a host instrument (not shown).
  • a host instrument not shown.
  • air pressure on either side of the liquid sample 9 is equal (see fig 2A).
  • the hemispherical bellow 8A is then actuated by pins present in the host instrument pressing down on its external surface and thus reducing the volume that air can fill between the bellow and the liquid sample.
  • the host instrument pins or actuators can be provided with sensors to detect the surface of the bellows to ensure specific control of the pressure is achieved.
  • the second bellow 8B is acting as a pressure 'reservoir' thus dampening some of the movement of the sample as the relative change in pressure is not as large, meaning the sample can still move, but pressure inside the channel can stay under 2 bar (but still increase compared to air pressure outside of the sealed cassette).
  • the absolute pressure inside the channel is increased to 1.6bar prior to the sample being amplified in the amplification zone.
  • figure 3 is a preferred variation of the present invention which shows that utilising a plurality of positive displacement pumps such as bellows or diaphragm pumps has additional benefits.
  • the preferred use of the present invention is to include at least two bellows pumps (or other positive displacement pumps) to induce an increase in pressure to at least 1.6 bar within the microchannel 2.
  • a liquid sample 9 is introduced into the inlet 5, and into the micro-fluidic channel 2; and the cap 6 is closed to form an air-tight seal.
  • the cassette 1 is placed into a host instrument (not shown). Prior to the cassette being closed by the cap 6, both of bellows pump 8A and bellows pump 8B are fully decompressed.
  • both first bellow 8A and second bellow 8B are actuated simultaneously to keep the overall volume of the system equal, and so keep the overall pressure of the system equal.
  • the host instrument applies a push force to the external surfaces of both first bellow 8A and second bellow 8B such that they are compressed by a first amount (the bellows are compressed approximately half-way) to increase the pressure inside the cassette to approx. 1.6 bar.
  • bellows pump 8A is positioned on the opposite side of the sample to bellows pump 8B i.e. a bellow is provided at either side of the sample, (in this case either side of the inlet 5 into which the sample was initially inserted) and in fact preferably at either side of the amplification zone.
  • the bellows pumps 8A and 8B can continue to be used to move the liquid sample 9 within the channel as required by the assay. By including various valves, and actuating them to open and close as required, this can allow sample to be very effectively moved throughout various regions of the micro- channel such that different actions and reactions can be carried out.
  • bellows pump 8A can be further compressed by a second amount and Bellow B allowed to re-inflate (the resilient material of the bellows allowing it to revert back to its original hemispherical shape).
  • Bellow B allowed to re-inflate (the resilient material of the bellows allowing it to revert back to its original hemispherical shape).
  • the pressure in the portion of the microchannel containing the amplification zone is increased to 1.6 bar prior to thermocycling occurring in the amplification zone to obtain the benefits of reducing bubble formation, as well as ensuring that the system works even at high altitude, whilst not damaging or exceeding the limits of any of the valves, caps or seals present in the cassette.
  • the pressure could also be raised to between 1.5bar and 2bar and significant beneficial effects would still be seen. In some embodiments even raising the pressure to l.lbar or 1.2bar or higher would still allow the benefits of the system working at higher altitudes and some reduction in bubble formation when compared to a standard system where no pressure increase prior to amplification would typically occur.

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Abstract

La présente invention concerne des dispositifs microfluidiques, tels que des cassettes pour le diagnostic, qui permettent un traitement sur cassette comprenant l'amplification par réaction en chaîne de la polymérase (PCR), et des systèmes et des procédés d'utilisation de ceux-ci, la cassette microfluidique étant mise sous pression avant l'amplification de l'acide nucléique dans ledit échantillon dans une section d'amplification/PCR du dispositif.
PCT/GB2019/053367 2018-11-29 2019-11-28 Dispositifs, systèmes et procédés microfluidiques améliorés Ceased WO2020109801A1 (fr)

Priority Applications (4)

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EP19816422.0A EP3887047A1 (fr) 2018-11-29 2019-11-28 Dispositifs, systèmes et procédés microfluidiques améliorés
US17/298,230 US20220088590A1 (en) 2018-11-29 2019-11-28 Improved microfluidic devices, systems and methods
JP2021531009A JP7499247B2 (ja) 2018-11-29 2019-11-28 改良されたマイクロ流体デバイス、システム及び方法
CN201980078467.9A CN113164955B (zh) 2018-11-29 2019-11-28 改进的微流体设备、系统和方法

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022084656A1 (fr) * 2020-10-19 2022-04-28 Quantumdx Group Limited Conditionnement thermique intégré et pcr dans un système de diagnostic poc moléculaire
JP2023042098A (ja) * 2021-09-14 2023-03-27 Nok株式会社 Ｐｃｒ用マイクロ流体チップおよびｐｃｒ分析装置

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN119307344B (zh) * 2023-07-11 2026-01-16 广州国家实验室 一种载体

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20090148933A1 (en) * 2006-03-15 2009-06-11 Micronics, Inc. Integrated nucleic acid assays
US20100129872A1 (en) * 2008-11-24 2010-05-27 Ghc Technologies, Inc. System and method for movement and positioning of reaction mixture during nucleic acid amplification
US20110039303A1 (en) * 2007-02-05 2011-02-17 Stevan Bogdan Jovanovich Microfluidic and nanofluidic devices, systems, and applications
US20180311673A1 (en) * 2016-01-05 2018-11-01 Nippon Sheet Glass Company, Limited Reaction processor, reaction processing vessel, and reaction processing method

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7763453B2 (en) * 2005-11-30 2010-07-27 Micronics, Inc. Microfluidic mixing and analytic apparatus

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20090148933A1 (en) * 2006-03-15 2009-06-11 Micronics, Inc. Integrated nucleic acid assays
US20110039303A1 (en) * 2007-02-05 2011-02-17 Stevan Bogdan Jovanovich Microfluidic and nanofluidic devices, systems, and applications
US20100129872A1 (en) * 2008-11-24 2010-05-27 Ghc Technologies, Inc. System and method for movement and positioning of reaction mixture during nucleic acid amplification
US20180311673A1 (en) * 2016-01-05 2018-11-01 Nippon Sheet Glass Company, Limited Reaction processor, reaction processing vessel, and reaction processing method

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022084656A1 (fr) * 2020-10-19 2022-04-28 Quantumdx Group Limited Conditionnement thermique intégré et pcr dans un système de diagnostic poc moléculaire
CN116419800A (zh) * 2020-10-19 2023-07-11 康特姆斯集团有限公司 分子poc诊断系统中的集成的热调节和pcr
JP2023546179A (ja) * 2020-10-19 2023-11-01 クァンタムディーエックス グループ リミテッド 分子ｐｏｃ診断システム内で統合された熱調節及びｐｃｒ
JP2023042098A (ja) * 2021-09-14 2023-03-27 Nok株式会社 Ｐｃｒ用マイクロ流体チップおよびｐｃｒ分析装置
JP7784257B2 (ja) 2021-09-14 2025-12-11 Nok株式会社 Ｐｃｒ用マイクロ流体チップおよびｐｃｒ分析装置

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CN113164955B (zh) 2023-06-09
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GB201819419D0 (en) 2019-01-16
US20220088590A1 (en) 2022-03-24
CN113164955A (zh) 2021-07-23

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